Life Sciences Commons^™

Comparison Of Human Urinary Exosomes Isolated Via Ultracentrifugation Alone Versus Ultracentrifugation Followed By Sec Column-Purification, Kun Huang, Sudha Garimella, Alyssa Clay-Gilmour, Lucia Vojtech, Bridget Armstrong, Madison Bessonny, Alexis Stamatikos

Faculty Publications

Chronic kidney disease is a progressive, incurable condition that involves a gradual loss of kidney function. While there are no non-invasive biomarkers available to determine whether individuals are susceptible to developing chronic kidney disease, small RNAs within urinary exosomes have recently emerged as a potential candidate to use for assessing renal function. Ultracentrifugation is the gold standard for urinary exosome isolation. However, extravesicular small RNA contamination can occur when isolating exosomes from biological fluids using ultracentrifugation, which may lead to misidentifying the presence of certain small RNA species in human urinary exosomes. Therefore, we characterized human urinary exosomal preparations isolated …

Mass Removal Efficiencies In Water And Consequences After A River Diversion Into Coastal Wetlands: Second Thoughts, R. Eugene Turner, Erik M. Swenson, James M. Lee, Charles S. Milan

Faculty Publications

Salinity control, nutrient additions, and sediment supply were directly or indirectly the rationale for a $220 million coastal wetland restoration project (Davis Pond River Diversion) that began in 2002. We sampled Mississippi River water going in and out of the receiving basin from 1999 to 2018 to understand why wetland loss increased after it began. There was a reduction in inorganic sediments, nitrogen (N), and phosphorus (P) concentrations within the ponding area of 77%, 39% and 34%, respectively, which is similar to that in other wetlands. But the average sediment accumulation of 0.6 mm year(-1) inadequately balances the present-day 5.6 …

Singing Strategies Are Linked To Perch Use On Foraging Territories In Heart-Nosed Bats, Grace C. Smarsh, Ashley M. Long, Michael Smotherman

Faculty Publications

Acoustic communication allows animals to coordinate and optimize resource utilization in space. Cardioderma cor, the heart-nosed bat, is one of the few species of bats known to sing during nighttime foraging. Previous research found that heart-nosed bats react aggressively to song playback, supporting the territorial defense hypothesis of singing in this species. We further investigated the territorial defense hypothesis from an ecological standpoint, which predicts that singing should be associated with exclusive areas containing a resource, by tracking 14 individuals nightly during the dry seasons in Tanzania. We quantified the singing behavior of individuals at all perches used throughout the …

Docking And Molecular Dynamic Of Microalgae Compounds As Potential Inhibitors Of Beta-Lactamase, Roberto Pestana-Nobles, Yani Aranguren-Diaz, Elwi Machado-Sierra, Juvenal Yosa, Nataly J. Galan-Freyle,, Laura X. Sepulveda-Montano, Daniel G. Kuroda, Leonardo C. Pacheco-Londono

Faculty Publications

Bacterial resistance is responsible for a wide variety of health problems, both in children and adults. The persistence of symptoms and infections are mainly treated with beta-lactam antibiotics. The increasing resistance to those antibiotics by bacterial pathogens generated the emergence of extended-spectrum beta-lactamases (ESBLs), an actual public health problem. This is due to rapid mutations of bacteria when exposed to antibiotics. In this case, beta-lactamases are enzymes used by bacteria to hydrolyze the beta-lactam rings present in the antibiotics. Therefore, it was necessary to explore novel molecules as potential beta-lactamases inhibitors to find antibacterial compounds against infection caused by ESBLs. …

Whole-Genome Sequencing And Rna-Seq Reveal Differences In Genetic Mechanism For Flowering Response Between Weedy Rice And Cultivated Rice, Richard S. Garcia, Sapphire Coronejo, Jonathan Concepcion, Prasanta K. Subudhi

Faculty Publications

Flowering is a key agronomic trait that influences adaptation and productivity. Previous studies have indicated the genetic complexity associated with the flowering response in a photoinsensitive weedy rice accession PSRR-1 despite the presence of a photosensitive allele of a key flowering gene Hd1. In this study, we used whole-genome and RNA sequencing data from both cultivated and weedy rice to add further insights. The de novo assembly of unaligned sequences predicted 225 genes, in which 45 were specific to PSRR-1, including two genes associated with flowering. Comparison of the variants in PSRR-1 with the 3K rice genome (RG) dataset identified …

Support Systems And Patient Care Delivery For Nonnative English-Speaking Patients: A Study Of Secondary School Athletic Trainers, Brea M. Stanton, Matthew J. Rivera, Zachary K. Winkelmann Phd, Lat, Atc, Lindsey E. Eberman

Faculty Publications

Context

Nonnative English speakers (NNESs) in the United States have more than doubled since 1990, increasing the likelihood of their seeking health care and experiencing language barriers. Language barriers in health care result in ineffective communication, a decreased level of care, and a reduction in overall provider satisfaction.

Objective

To investigate the experiences of secondary school athletic trainers (ATs) who provided care to patients who were NNESs or communicated with their NNES support systems.

Design

Qualitative study.

Setting

Semistructured interviews.

Patients or Other Participants

Fifteen secondary school ATs with experience communicating with NNES patients or their support systems.

Data Collection …

Offshore Spreading Of A Supercritical Plume Under Upwelling Wind Forcing: A Case Study Of The Winyah Bay Outflow, Alexander E. Yankovsky, Douglas Cahl, George Voulgaris

Faculty Publications

In this study, we present observations of the Winyah Bay (WB) plume (SC, United States) formed by high freshwater discharge and a moderate upwelling-favorable wind acting continuously for ∼1.5 days prior to the shipboard survey. If a similar wind forcing persists over a longer period, the plume turns upstream (against its natural propagation) and curves offshore forming a “filament” with minimal transverse spreading, as seen in numerous satellite images. The observed plume comprises a train of tidal sub-plumes undergoing rotational adjustment and being transported offshore by Ekman dynamics. The WB outflow is supercritical in terms of the interior Froude number. …

Reply To Comment: Controls On Turnover Of Marine Dissolved Organic Matter-Testing The Null Hypothesis Of Purely Concentration-Driven Uptake, Yuan Shen, Ronald Benner

Faculty Publications

Our recent bioassay experiments indicate that molecular properties are a primary control on the microbial utilization of dissolved organic matter in the ocean. This finding is questioned by Lennartz and Dittmar who modeled our experiments and concluded that our observations could be largely explained by concentration-driven uptake independent from molecular properties. We suggest the authors' models are deficient for establishing the relative roles of molecular properties and concentration-driven uptake. Our conclusion is consistent with earlier and recent experimental results and biogeochemical observations, supporting a unified theory with molecular properties as a more prominent control than concentration-driven uptake on marine organic …

Transcriptional Coactivators: Driving Force Of Plant Immunity, Muhammad Saad Shoaib Khan, Faisal Islam, Huan Chen, Ming Chang, Daowen Wang, Fengquan Liu, Zhengqing Fu, Jian Chen

Faculty Publications

Salicylic acid (SA) is a plant defense signal that mediates local and systemic immune responses against pathogen invasion. However, the underlying mechanism of SA-mediated defense is very complex due to the involvement of various positive and negative regulators to fine-tune its signaling in diverse pathosystems. Upon pathogen infections, elevated level of SA promotes massive transcriptional reprogramming in which Non-expresser of PR genes 1 (NPR1) acts as a central hub and transcriptional coactivator in defense responses. Recent findings show that Enhanced Disease Susceptibility 1 (EDS1) also functions as a transcriptional coactivator and stimulates the expression of PR1 in the presence of …

Examination Of The Prevalence Of Female Athlete Triad Components Among Competitive Cheerleaders, Allison B. Smith, Jennifer L. Gay, Shawn M. Arent Phd, Cscs*D, Fissn, Facsm, Fnak, Mark A. Sarzynski Ph.D., Faha, Facsm, Dawn M. Emerson, Toni Torres-Mcgehee Phd, Lat, Atc

Faculty Publications

The purpose of this study was to examine individual and combined Female Athlete Triad components within collegiate cheerleaders, an at-risk group. Cheerleaders ( = 19; age: 20.3 ± 1.2 years) completed anthropometric measurements, health history questionnaires, resting metabolic rate, the eating disorder inventory-3 and symptom checklist, blood sample, and DXA scan. Participants completed dietary and exercise logs for 7 days and used heart rate monitors to track daily and exercise energy expenditure. Proportions were calculated for low energy availability (LEA) risk, disordered eating risk, and pathogenic behaviors. Chi-square analysis was used to determine the difference between cheerleaders who experience low …

An Exploratory Study On The Microbiome Of Northern And Southern Populations Of Ixodes Scapularis Ticks Predicts Changes And Unique Bacterial Interactions, Deepak Kumar, Latoyia P. Downs, Abdulsalam Adegoke, Erika Machtinger, Kelly Oggenfuss, Richard S. Ostfeld, Monica Embers, Shahid Karim

Faculty Publications

The black-legged tick (Ixodes scapularis) is the primary vector of Borrelia burgdorferi, the causative agent of Lyme disease in North America. However, the prevalence of Lyme borreliosis is clustered around the Northern States of the United States of America. This study utilized a metagenomic sequencing approach to compare the microbial communities residing within Ix. scapularis populations from northern and southern geographic locations in the USA. Using a SparCC network construction model, we performed potential interactions between members of the microbial communities from Borrelia burgdorferi–infected tissues of unfed and blood-fed ticks. A significant difference in bacterial composition …

Reef Sediments Can Act As A Stony Coral Tissue Loss Disease Vector, Michael S. Studivan, Ashley M. Rossin, Ewelina Rubin, Nash Soderberg, Daniel M. Holstein, Ian C. Enochs

Faculty Publications

Stony coral tissue loss disease (SCTLD) was first observed in 2014 near Virginia Key in Miami-Dade County, Florida. Field sampling, lab experiments, and modeling approaches have suggested that reef sediments may play a role in SCTLD transmission, though a positive link has not been tested experimentally. We conducted an ex situ transmission assay using a statistically-independent disease apparatus to test whether reef sediments can transmit SCTLD in the absence of direct contact between diseased and healthy coral tissue. We evaluated two methods of sediment inoculation: batch inoculation of sediments collected from southeast Florida using whole colonies of diseased Montastraea cavernosa, …

Improved Chromosome-Level Genome Assembly Of The Glanville Fritillary Butterfly (Melitaea Cinxia) Integrating Pacific Biosciences Long Reads And A High-Density Linkage Map, Olli-Pekka Smolander, Daniel Blande, Virpi Ahola, Pasi Rastas, Jaakko Tanskanen, Juhana Kammonen, Vicencio Oostra, Lorenzo Pellegrini, Suvi Ikonen, Tad Dallas, Michelle F. Dileo, Anne Duplouy, Iihan Cem Duru, Pauliina Halimaa, Aapo Kahilainen, Suyog S. Kuwar, Sirpa O. Kärenlampi, Elvira Lafuente, Shiqi Luo, Jenny Makkonen, Abhilash Nair, Narua De La Paz Celorio-Mancera, Ville Pennanen, Annukka Ruokolainen, Tarja Sundell, Arja I. Tervahauta, Victoria Twort, Erik Van Bergen, Janina Österman-Udd, Lars Paulin, Mikko J. Frilander, Petri Auvinen, Marjo Saastamoinen

Faculty Publications

Background

The Glanville fritillary (Melitaea cinxia) butterfly is a model system for metapopulation dynamics research in fragmented landscapes. Here, we provide a chromosome-level assembly of the butterfly's genome produced from Pacific Biosciences sequencing of a pool of males, combined with a linkage map from population crosses.

Results

The final assembly size of 484 Mb is an increase of 94 Mb on the previously published genome. Estimation of the completeness of the genome with BUSCO indicates that the genome contains 92–94% of the BUSCO genes in complete and single copies. We predicted 14,810 genes using the MAKER pipeline and …

Evolutionary Analyses Of Visual Opsin Genes In Frogs And Toads: Diversity, Duplication, And Positive Selection, Ryan K. Schott, Leah Perez, Matthew Kwiatkowski, Vance Imhoff, Jennifer M. Gumm

Faculty Publications

Among major vertebrate groups, anurans (frogs and toads) are understudied with regard to their visual systems, and little is known about variation among species that differ in ecology. We sampled North American anurans representing diverse evolutionary and life histories that likely possess visual systems adapted to meet different ecological needs. Using standard molecular techniques, visual opsin genes, which encode the protein component of visual pigments, were obtained from anuran retinas. Additionally, we extracted the visual opsins from publicly available genome and transcriptome assemblies, further increasing the phylogenetic and ecological diversity of our dataset to 33 species in total. We found …

Chronic Background Radiation Correlates With Sperm Swimming Endurance In Bank Voles From Chernobyl, Kati Kivisaari, Sara Calhim, Zbyszek Boratyríski, Timothy Mousseau, Anders P. Møller, Tapio Mappes

Faculty Publications

Sperm quantity and quality are key features explaining intra- and interspecific variation in male reproductive success. Spermatogenesis is sensitive to ionizing radiation and laboratory studies investigating acute effects of ionizing radiation have indeed found negative effects of radiation on sperm quantity and quality. In nature, levels of natural background radiation vary dramatically, and chronic effects of low-level background radiation exposure on spermatogenesis are poorly understood. The Chernobyl region offers a unique research opportunity for investigating effects of chronic low-level ionizing radiation on reproductive properties of wild organisms. We captured male bank voles (Myodes glareolus) from 24 locations in the Chernobyl …

Investigation Of Metastable Low Dimensional Halometallates, Navindra Keerthisinghe, Matthew S. Christian, Anna A. Berseneva, Gregory Morrison, Vladislav V. Klepov, Mark D. Smith, Hans Conrad Zur Loye

Faculty Publications

The solvothermal synthesis, structure determination and optical characterization of five new metastable halometallate compounds, [1,10-phenH][Pb_3.5I₈] (1), [1,10-phenH₂][Pb₅I₁₂]·(H₂O) (2), [1,10-phen][Pb₂I₄] (3), [1,10-phen]₂[Pb₅Br₁₀] (4) and [1,10-phenH][SbI₄]·(H₂O) (5), are reported. The materials exhibit rich structural diversity and exhibit structural dimensionalities that include 1D chains, 2D sheets and 3D frameworks. The optical spectra of these materials are consistent with bandgaps ranging from 2.70 to 3.44 eV. We show …

Electrophysiological Responses To Conspecific Odorants In Xenopus Laevis Show Potential For Chemical Signaling, Heather J. Rhodes, Melanie Amo

Faculty Publications

No abstract provided.

A Qualitative Report Of The Perceptions Of The Covid-19 Pandemic From Collegiate Student-Athletes, Dylan C. Rowe, Zachary K. Winkelmann, Shawn M. Arent, Michelle A. Arent, Alexa J. Chandler, Nancy A. Uriegas, Toni M. Torres-Mcgehee

Faculty Publications

CONTEXT: The COVID-19 pandemic led to an abrupt disruption in access to services and personnel for collegiate student-athletes in the spring and summer of 2020. We sought to identify the effects of this unprecedented change by examining the psychological well-being, changes to normal routines, and return-to-play considerations of current student-athletes in order to guide support for both current and future student-athletes who may face similar situations. METHODS: We utilized a phenomenological approach to interview a purposeful sample of eighteen collegiate student-athletes (7 males, 11 females; mean age = 20 years) from across the United States. The participants were interviewed using …

Factors Related To Species Richness, Endemism, And Conservation Status Of The Herpetofauna (Amphibia And Reptilia) Of Mexican States, Geoffrey R. Smith, Julio A. Lemos-Espinal

Faculty Publications

No abstract provided.

Structure Of A Monomeric Photosystem Ii Core Complex From A Cyanobacterium Acclimated To Far-Red Light Reveals The Functions Of Chlorophylls D And F, Christopher J. Gisriel, Gaozhong Shen, Ming-Yang Ho, Vasily Kurashov, David A. Flesher

Faculty Publications

Far-red light (FRL) photoacclimation in cyanobacteria provides a selective growth advantage for some terrestrial cyanobacteria by expanding the range of photosynthetically active radiation to include far-red/near-infrared light (700-800 nm). During this photoacclimation process, photosystem II (PSII), the water:plastoquinone photooxidoreductase involved in oxygenic photosynthesis, is modified. The resulting FRL-PSII is comprised of FRL-specific core subunits and binds chlorophyll (Chl) d and Chl f molecules in place of several of the Chl a molecules found when cells are grown in visible light. These new Chls effectively lower the energy canonically thought to define the "red limit" for light required to drive photochemical …

On The Need For New Measures Of Phylogenomic Support, Robert C. Thomson, Jeremy M. Brown

Faculty Publications

The scale of data sets used to infer phylogenies has grown dramatically in the last decades, providing researchers with an enormous amount of information with which to draw inferences about evolutionary history. However, standard approaches to assessing confidence in those inferences (e.g., nonparametric bootstrap proportions [BP] and Bayesian posterior probabilities [PPs]) are still deeply influenced by statistical procedures and frameworks that were developed when information was much more limited. These approaches largely quantify uncertainty caused by limited amounts of data, which is often vanishingly small with modern, genome-scale sequence data sets. As a consequence, today's phylogenomic studies routinely report near-complete …

Analyzing Factors Influencing Secondary Agricultural Education Teachers’ Skill Level In Welding, Rafael Bakhtavoryan, Douglas D. Lavergne

Faculty Publications

The effects of secondary agricultural education teachers’ demographic characteristics on their propensity to become more skilled in welding were examined by estimating an ordered logit model for teachers in Northeast Texas. The sample of teachers was derived from the Agricultural Mechanic Competency Development survey for 2015. According to the estimation results, relative to female, being male was positively associated with being more skilled in welding. Compared to having a Master’s degree, having only a Bachelor’s degree was negatively associated with being more skilled in welding. Having five or less years of teaching positively affected becoming more skilled in welding compared …

The Effect Of Wearable Activity Monitor Presence On Step Counts, William B. Tayler, James D. Lecheminant, Jospeh Price, Christian P. Tadje

Faculty Publications

Objective: In this study, we tested the effect of wearable activity monitors and tracking on physical activity over 2 weeks.

Methods: Ninety young adults participated. Prior to initiation of the study, each participant was asked to download an iPhone app that tracked physical activity level (step counts) but were not told the app’s purpose. Each participant was then randomly assigned to one of 3 groups (N=30/group): (1) measurement awareness, (2) measurement awareness+track/record, or (3) control. Participants in the measurement awareness and the measurement awareness+track/record groups were given a pedometer to wear for 2 weeks. Additionally, participants in the measurement awareness+track/ …

Hydrodynamic Assessment Of Natural And Nature-Based Features For Escatawpa River And Grand Bay In The Northern Gulf Of Mexico, Peter Bacopoulos, Karim Alizad, Davina Passeri, Matthew Bilskie, Stephen Medeiros, Scott Hagen

Faculty Publications

This presentation showcases a hydrodynamic assessment of natural and nature-based features (NNBFs) for the Pascagoula River, the Escatawpa River and Grand Bay, located along the Mississippi coast of the northern Gulf of Mexico. Two separate NNBF projects are being considered to: (1) restore the historical footprint (ca. 1848) of Grand Batture Island for coastal protection purposes; and (2) reconnect the hydraulics between the Escatawpa River and Grand Bay for ecosystem services purposes. The intended coastal protection benefits of the first project include buffering agency to wave attack and attenuation of storm surge with the restored island. The intended ecosystem services …

Single-Variable Porous Nanomaterial Series From Polymer Structure-Directing Agents, Morgan Stefik

Faculty Publications

Block polymer structure-directing agents (SDA) enable the production of porous nanoscale materials. Most strategies rely upon polymer equilibration where diverse morphologies are realized in porous functional materials. This review details how solvent selectivity determines the polymer SDA behaviors, spanning from bulk-type to solution-type. Equilibrating behavior of either type, however, obscures nanostructure cause-and-effect since the resulting sample series convolve multiple spatial variations. Solution-type SDA behaviors include both dynamic and persistent micelles. Persistent micelle templates (PMT) use high solvent selectivity for kinetic entrapment. PMTs enable independent wall thickness control with demonstrated 2 Å precision alterations. Unimodal PMT pore size distributions have spanned …

Genomics And Transcriptomics Landscapes Associated To Changes In Insulin Sensitivity In Response To Endurance Exercise Training, Louise Y. Takeshita, Peter K. Davidsen, John M. Herbert, Philipp Antczak, Matthijs K.C. Hesslink, Patrick Schrauwen, S. John Weisnagel, Jeremy M. Robbins, Robert E. Gerszten, Sujoy Ghosh, Mark A. Sarzynski Ph.D., Faha, Facsm, Claude Bouchard, Francesco Falciani

Faculty Publications

Despite good adherence to supervised endurance exercise training (EET), some individuals experience no or little improvement in peripheral insulin sensitivity. The genetic and molecular mechanisms underlying this phenomenon are currently not understood. By investigating genome-wide variants associated with baseline and exercise-induced changes (∆) in insulin sensitivity index (S_i) in healthy volunteers, we have identified novel candidate genes whose mouse knockouts phenotypes were consistent with a causative effect on S_i. An integrative analysis of functional genomic and transcriptomic profiles suggests genetic variants have an aggregate effect on baseline S_i and ∆S_i, focused around cholinergic …

International Society Of Sports Nutrition Position Stand: Caffeine And Exercise Performance, Nanci S. Guest, Trisha A. Vandusseldorp, Michael T. Nelson, Jozo Grgic, Brad J. Schoenfeld, Nathaniel D.M. Jenkins, Shawn M. Arent Ph.D., Cscs*D, Fissn, Facsm, Fnak, Jose Antonio, Jeffrey R. Stout, Eric T. Trexler, Abbie E. Smith-Ryan, Erica R. Goldstein, Douglas S. Kalman, Bill I. Campbell

Faculty Publications

Following critical evaluation of the available literature to date, The International Society of Sports Nutrition (ISSN) position regarding caffeine intake is as follows:1.Supplementation with caffeine has been shown to acutely enhance various aspects of exercise performance in many but not all studies. Small to moderate benefits of caffeine use include, but are not limited to: muscular endurance, movement velocity and muscular strength, sprinting, jumping, and throwing performance, as well as a wide range of aerobic and anaerobic sport-specific actions.2.Aerobic endurance appears to be the form of exercise with the most consistent moderate-to-large benefits from caffeine use, although the magnitude of …

Environmental Rnai Pathways In The Two-Spotted Spider Mite, Mosharrof Mondal, Jacob Peter, Obrie Scarbrough, Alex Flynt

Faculty Publications

© 2020, The Author(s).

Background:RNA interference (RNAi) regulates gene expression in most multicellular organisms through binding of small RNA effectors to target transcripts. Exploiting this process is a popular strategy for genetic manipulation and has applications that includes arthropod pest control. RNAi technologies are dependent on delivery method with the most convenient likely being feeding, which is effective in some animals while others are insensitive. The two-spotted spider mite, Tetranychus urticae, is prime candidate for developing RNAi approaches due to frequent occurrence of conventional pesticide resistance. Using a sequencing-based approach, the fate of ingested RNAs was explored to …

Spectroscopic Studies Of The Interaction Between Phosphorus Heterocycles And Cytochrome P450, Dumei Ma, Libo Zhang, Yingwu Yin, Yuxing Gao, Qian Wang

Faculty Publications

P450 fatty acid decarboxylase OleT from Staphylococcus aureus (OleT_SA) is a novel cytochrome P450 enzyme that catalyzes the oxidative decarboxylation of fatty acids to yield primarily terminal alkenes and CO₂ or minor α- and β-hydroxylated fatty acids as side-products. In this work, the interactions between a series of cycloalkyl phosphorus heterocycles (CPHs) and OleT_SA were investigated in detail by fluorescence titration experiment, ultraviolet–visible (UV–vis) and ³¹P NMR spectroscopies. Fluorescence titration experiment results clearly showed that a dynamic quenching occurred when CPH-6, a representative CPHs, interacted with OleT_SA with a binding constant value of 15.2 …

An Acute Naproxen Dose Does Not Affect Core Temperature Or Interleukin-6 During Cycling In A Hot Environment, Dawn M. Emerson, Stephen Cl Chen, Toni M. Torres-Mcgehee, Craig E. Pfeifer, Charles C. Emerson, J Mark Davis

Faculty Publications

Non-steroidal anti-inflammatory drugs' anti-pyretic and anti-inflammatory effects has led some individuals to theorize these medications may blunt core body temperature (Tc) increases during exercise. We utilized a double-blind, randomized, and counterbalanced cross-over design to examine the effects of a 24-h naproxen dose (3-220 mg naproxen pills) and placebo (0 mg naproxen) on Tc and plasma interleukin-6 (IL-6) concentrations during cycling in a hot or ambient environment. Participants ( = 11; 6 male, 5 female; age = 27.8 ± 6.5 years, weight = 79.1 ± 17.9 kg, height = 177 ± 9.5 cm) completed 4 conditions: 1) placebo and ambient (Control); …